CN110964690A - Method for preventing gamete mismatching in implementation process of assisted reproduction technology - Google Patents
Method for preventing gamete mismatching in implementation process of assisted reproduction technology Download PDFInfo
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- CN110964690A CN110964690A CN201911321615.3A CN201911321615A CN110964690A CN 110964690 A CN110964690 A CN 110964690A CN 201911321615 A CN201911321615 A CN 201911321615A CN 110964690 A CN110964690 A CN 110964690A
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- blastocyst
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- paternity
- ovum
- free dna
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- 238000000034 method Methods 0.000 title claims abstract description 27
- 230000008569 process Effects 0.000 title claims abstract description 13
- 210000002459 blastocyst Anatomy 0.000 claims abstract description 71
- 210000004027 cell Anatomy 0.000 claims abstract description 39
- 210000004681 ovum Anatomy 0.000 claims abstract description 18
- 108010000912 Egg Proteins Proteins 0.000 claims abstract description 15
- 102000002322 Egg Proteins Human genes 0.000 claims abstract description 15
- 210000001161 mammalian embryo Anatomy 0.000 claims abstract description 13
- 238000012360 testing method Methods 0.000 claims abstract description 13
- 230000002068 genetic effect Effects 0.000 claims abstract description 12
- 239000002773 nucleotide Substances 0.000 claims abstract description 7
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 7
- 230000004720 fertilization Effects 0.000 claims abstract description 6
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 6
- 238000012163 sequencing technique Methods 0.000 claims abstract description 6
- 238000012546 transfer Methods 0.000 claims abstract description 6
- 230000003321 amplification Effects 0.000 claims description 3
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 3
- 238000001514 detection method Methods 0.000 abstract description 5
- 238000011161 development Methods 0.000 abstract description 4
- 239000000463 material Substances 0.000 abstract description 2
- 235000013601 eggs Nutrition 0.000 description 10
- 238000012258 culturing Methods 0.000 description 4
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 210000002257 embryonic structure Anatomy 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 210000003754 fetus Anatomy 0.000 description 1
- 230000008774 maternal effect Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000008775 paternal effect Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0603—Embryonic cells ; Embryoid bodies
- C12N5/0604—Whole embryos; Culture medium therefor
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B17/00—Surgical instruments, devices or methods, e.g. tourniquets
- A61B17/42—Gynaecological or obstetrical instruments or methods
- A61B17/425—Gynaecological or obstetrical instruments or methods for reproduction or fertilisation
- A61B17/43—Gynaecological or obstetrical instruments or methods for reproduction or fertilisation for artificial insemination
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B17/00—Surgical instruments, devices or methods, e.g. tourniquets
- A61B17/42—Gynaecological or obstetrical instruments or methods
- A61B17/425—Gynaecological or obstetrical instruments or methods for reproduction or fertilisation
- A61B17/435—Gynaecological or obstetrical instruments or methods for reproduction or fertilisation for embryo or ova transplantation
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6869—Methods for sequencing
Abstract
The invention is applicable to the technical field of assisted reproduction. The invention discloses a method for preventing gamete mismatch in the implementation process of an assisted reproduction technology, which comprises the steps of combining an ovum of a receptor with a sperm approved by the receptor to finish ovum fertilization, then placing the fertilized ovum in a culture solution to be cultured to a blastocyst, extracting blastocyst cell free DNA from the blastocyst culture solution, analyzing and obtaining Single Nucleotide Polymorphism (SNP) sites by using a gene sequencing technology to perform affinity genetic identification on the blastocyst cells, and transplanting the blastocyst to the receptor when the sites and the receptor accord with a paternity relationship; embryo transfer is terminated when the site is not in paternity with the recipient. Because the blastocyst cell free DNA is selected to carry out the biological genetic paternity test on the blastocyst before the blastocyst is transplanted, reliable biological detection materials are obtained without destroying the tissue structure of the blastocyst to carry out paternity test, the growth and development of the blastocyst are not influenced, and meanwhile, the blastocyst and a receptor can have a determined biological genetic paternity relationship, thereby avoiding causing serious consequences.
Description
Technical Field
The invention relates to the technical field of assisted reproduction, in particular to a method for preventing gamete mismatching in the implementation process of an assisted reproduction technology.
Background
With the development of artificial assisted reproduction technology, more and more couples incapable of breeding realize the desire of breeding. Assisted reproduction is generally achieved by transferring the embryo in vitro into the body, and when the embryo survives and grows normally in vivo, the embryo can grow normally. Because many auxiliary reproduction objects managed by an artificial auxiliary reproduction mechanism are managed, the processes of taking eggs, fertilizing sperm and eggs in vitro, culturing embryos in the early stage, transferring embryos and the like in the auxiliary reproduction process are complex, time intervals exist among all the steps, and the collection of sperms and the collection of eggs are both manual operation, so that errors are difficult to avoid when a plurality of collection objects are collected, the collection of sperms and eggs cannot be detected nondestructively, the detection can be usually carried out only when the fertilized eggs are transplanted in vivo, fetal DNA is obtained through a puncture operation for detection, and serious consequences are produced when the non-paternity relationship between the fetus and a receptor is detected.
Disclosure of Invention
The invention mainly solves the technical problem of providing a method for preventing gamete mismatching in the implementation process of an assisted reproduction technology, and the method for preventing gamete mismatching in the implementation process of the assisted reproduction technology can avoid the non-parental condition of an embryo and a receptor before embryo transfer.
In order to solve the technical problems, the invention provides a method for preventing gamete mismatch in the implementation process of an assisted reproduction technology, which comprises the steps of combining an ovum of a receptor with a sperm approved by the receptor to finish ovum fertilization, placing the fertilized ovum in a culture solution to be cultured to a blastocyst stage, extracting blastocyst cell free DNA from the blastocyst culture solution, analyzing and obtaining a single nucleotide polymorphism site by using a gene sequencing technology to carry out noninvasive genetic paternity test on the blastocyst cell, and transplanting the blastocyst to the receptor when the site conforms to the paternity; embryo transfer is terminated when the site does not correspond paternity to the recipient.
Further, when the blastocyst develops to 8-16 blastocyst cells, blastocyst cell free DNA is extracted for paternity test.
Further, the extracted blastocyst cell-free DNA is subjected to an amplification step prior to paternity testing of the blastocyst cells.
The invention relates to a method for preventing gamete mismatch in the implementation process of an assisted reproduction technology, which comprises the steps of combining an ovum of a receptor with a sperm approved by the receptor to finish ovum fertilization, then placing the fertilized ovum in a culture solution to be cultured to a blastocyst stage, extracting blastocyst cell free DNA from the blastocyst culture solution, analyzing and obtaining a single nucleotide polymorphism site by using a gene sequencing technology to carry out noninvasive genetic paternity identification on the blastocyst cell, and transplanting the blastocyst to the receptor when the site conforms to the paternity; embryo transfer is terminated when the site does not correspond paternity to the recipient. Because the paternity test is not carried out on the cell embryo before the cell embryo is transplanted, the growth and development of the cell embryo are not influenced, and meanwhile, the serious consequences caused by no paternity relationship between the cell embryo and a receptor can be avoided.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the description are some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on the drawings without creative efforts.
Fig. 1 is a flow chart of an embodiment of a method for preventing gamete mismatch in the implementation process of an assisted reproduction technology.
The objectives, features, and advantages of the present invention will be further explained with reference to the accompanying drawings.
Detailed Description
The following claims of the present invention are further detailed in conjunction with the detailed description of the embodiments and the accompanying drawings, and it is apparent that the embodiments described are only a part of the embodiments of the present invention, and are all embodiments. All other embodiments obtained by persons of ordinary skill in the art based on the embodiments of the present invention without any inventive work also belong to the protection scope of the present invention.
It should be understood that in the description of the present invention, all directional terms such as "upper", "lower", "left", "right", "front", "rear", etc., indicate orientations or positional relationships that are based on the orientations or positional relationships shown in the drawings or the orientations or positional relationships in which the products of the present invention are conventionally placed when in use, are used only for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the devices or elements being referred to must have a specific orientation, be constructed in a specific orientation, and be operated. The directional indication may be changed when the specific posture is changed, and thus, the present invention is not to be construed as limited.
Furthermore, ordinal words such as "first", "second", etc., are used for differentiation only and are not to be construed as indicating or implying any relative importance or implicit indication of the number of technical features indicated. The technical features "first" and "second" may be explicit or implicit and at least one of the technical features may be limited thereby. In the description of the present invention, "a plurality" means at least two, i.e., two or more, unless expressly defined otherwise; the meaning of "at least one" is one or more than one.
As shown in fig. 1, the present invention provides an embodiment of a method for preventing gamete mismatch during implementation of assisted reproduction technology.
The method for preventing gamete mismatch in the implementation process of the assisted reproduction technology comprises the following steps:
and step S10, fertilizing the ovum, specifically, combining the acceptor ovum with the acceptor approved sperm to finish the fertilization of the ovum, wherein the acceptor approved sperm firstly meets the legal requirements and secondly accepts the acceptor sperm.
And step S11, culturing the fertilized egg, specifically, placing the fertilized egg in the culture solution in the step S10 to be cultured to a blastocyst stage.
And step S12, extracting blastocyst cell free DNA, specifically, extracting blastocyst cell free DNA from a blastocyst culture solution when the blastocyst develops to 8-16 blastocyst cells.
Step S13, paternity test is carried out on the extracted blastocyst cell free DNA, specifically, the blastocyst cell free DNA is analyzed by utilizing gene sequencing technology, and the Single Nucleotide Polymorphism (SNP) site of the blastocyst cell is obtained to carry out noninvasive biogenetic paternity test on the blastocyst cell.
And step S14, when the sites and the receptors detected in the step S13 do not conform to the biological genetic parent-child relationship, terminating blastocyst culture or blastocyst transfer.
And step S15, when the sites detected in the step S13 and the receptors accord with the biological genetic parent-child relationship, continuing to culture blastocysts or transplanting the blastocysts.
Specifically, the collection of ova and sperms, the fertilization of ova, the culture of fertilized ova, the extraction of blastocyst cell free DNA, and the analysis of blastocyst cell free DNA by using a gene sequencing technology to obtain Single Nucleotide Polymorphism (SNP) sites of the blastocyst cells and perform genetic paternity relationship identification on the blastocyst cells are realized by adopting the prior art and are not the main points of the invention.
And placing the fertilized eggs in a culture solution for culturing, wherein the culture environment and the culture solution of the fertilized eggs are conventional fertilized eggs. And extracting blastocyst cell free DNA when the blastocyst develops to 8-16 blastocyst cells, and performing amplification to increase the number of the blastocyst cell free DNA, so that the detection is easier. Extracting blastocyst cell free DNA for paternity test, identifying the blastocyst cell paternity by acquiring Single Nucleotide Polymorphism (SNP) sites of the blastocyst cells, and continuously culturing the blastocyst cells until the blastocyst cells are transplanted to a receptor when the sites and the receptor accord with the paternity relationship; blastocyst cell culture is terminated when the site does not correspond paternity to the receptor.
Because the blastocyst cell free DNA is selected to carry out the biological genetic paternity test on the blastocyst before the blastocyst is transplanted, reliable biological detection materials are obtained without destroying the tissue structure of the blastocyst to carry out the paternity test, the growth and the development of the blastocyst are not influenced, and meanwhile, the blastocyst and the receptor can have a determined biological genetic paternity relationship, so that the serious consequences caused by the non-maternal and paternal biological genetic paternity relationship between the blastocyst and the receptor are avoided.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: it is to be understood that modifications may be made to the above-described arrangements in the embodiments or equivalents may be substituted for some of the features of the embodiments described above, and such modifications or substitutions may be made without departing from the spirit and scope of the present invention in its aspects.
Claims (3)
1. The method for preventing gamete mismatch in the implementation process of the assisted reproduction technology comprises the steps of combining an ovum of a receptor with a sperm approved by the receptor to finish ovum fertilization, placing the fertilized ovum in a culture solution to be cultured to a blastocyst stage, extracting blastocyst cell free DNA from the blastocyst culture solution, analyzing and obtaining a single nucleotide polymorphism site by using a gene sequencing technology to carry out noninvasive genetic paternity identification on the blastocyst cell, and transplanting the blastocyst to the receptor when the site and the receptor accord with the paternity; embryo transfer is terminated when the site does not correspond paternity to the recipient.
2. The method for preventing gamete mismatch during implementation of assisted reproduction technology of claim 1, wherein blastocyst cell free DNA is extracted for paternity testing when the blastocyst develops to 8-16 blastocyst cells.
3. The method of claim 1, wherein the amplification step is performed on the extracted blastocyst cell-free DNA prior to paternity testing of the blastocyst cells.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2019100409317 | 2019-01-16 | ||
| CN201910040931 | 2019-01-16 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110964690A true CN110964690A (en) | 2020-04-07 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911321615.3A Pending CN110964690A (en) | 2019-01-16 | 2019-12-20 | Method for preventing gamete mismatching in implementation process of assisted reproduction technology |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120122701A1 (en) * | 2010-05-18 | 2012-05-17 | Gene Security Network, Inc. | Methods for Non-Invasive Prenatal Paternity Testing |
| CN105368936A (en) * | 2015-11-05 | 2016-03-02 | 上海序康医疗科技有限公司 | Method for detecting embryo chromosome abnormality by utilizing blastula culture solution |
| US20170002414A1 (en) * | 2014-01-30 | 2017-01-05 | Pécsi Tudományegyetem | Preimplantation assessment of embryos through detection of free embryonic dna |
| CN106399535A (en) * | 2016-10-19 | 2017-02-15 | 江苏苏博生物医学股份有限公司 | Method for detecting noninvasive paternity tests through high-throughput sequencing |
-
2019
- 2019-12-20 CN CN201911321615.3A patent/CN110964690A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120122701A1 (en) * | 2010-05-18 | 2012-05-17 | Gene Security Network, Inc. | Methods for Non-Invasive Prenatal Paternity Testing |
| US20170002414A1 (en) * | 2014-01-30 | 2017-01-05 | Pécsi Tudományegyetem | Preimplantation assessment of embryos through detection of free embryonic dna |
| CN105368936A (en) * | 2015-11-05 | 2016-03-02 | 上海序康医疗科技有限公司 | Method for detecting embryo chromosome abnormality by utilizing blastula culture solution |
| CN106399535A (en) * | 2016-10-19 | 2017-02-15 | 江苏苏博生物医学股份有限公司 | Method for detecting noninvasive paternity tests through high-throughput sequencing |
Non-Patent Citations (2)
| Title |
|---|
| 唐双柏等: "无创伤性产前诊断方法及其 法医学应用前景" * |
| 李刚;刘艳;孙莹璞;: "单核苷酸多态性微阵列在胚胎植入前遗传学诊断中的应用" * |
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